Antibiotic TA (TA) is a wide-spectrum, bactericidal antibiotic produced by Myxococcus xanthus str... more Antibiotic TA (TA) is a wide-spectrum, bactericidal antibiotic produced by Myxococcus xanthus strain TA. It was previously demonstrated that TA binds tightly to soft tissues while retaining its bactericidal activity in the bound form. The present study was undertaken to investigate TA adsorption to dental hard tissues. Slabs of dental tissues that had been cut from periodontally-involved extracted human teeth were treated with TA and then washed in saline with shaking (saline being replaced every 15 minutes). After 30, 60, and 120 minutes of washing, 45, 39, and 27% of the input TA activities were retained on the slabs, respectively. The tooth-bound TA was released slowly into the aqueous medium in active form. Similar experiments with beta-lactam antibiotics resulted in no significant adhesion to the slabs. The study demonstrates that TA binds tightly to dental tissues while retaining its bactericidal properties.
Bdellovibrios capable of axenic growth grow in a cell-free medium at a rate considerably lower th... more Bdellovibrios capable of axenic growth grow in a cell-free medium at a rate considerably lower than that attainable in a two-membered culture with Escherichia coli. The axenic growth rate may be improved either by adjustment of the osmosity of the medium or by the addition of low concentrations of spermine.
Autocide AMV of Myxococcus xanthus was purified and identified as phosphatidylethanolamine. Alkal... more Autocide AMV of Myxococcus xanthus was purified and identified as phosphatidylethanolamine. Alkaline hydrolysis of AMV yielded a high proportion of mono- and diunsaturated fatty acids. The bactericidal activity of AMV on M. xanthus depended upon the density of target cells: the greater the cell density, the greater the killing by AMV. For example, at 2 U of AMV per ml, 0, 50, and 99% killing was measured with 2 X 10(4), 2 X 10(5), and 2 X 10(7) target cells per ml, respectively. The cell-density-dependent activity of AMV was also observed on solid medium. Studies with model lipid compounds suggest that the inhibitory activity of AMV is due to the fatty acid moiety, released from phosphatidylethanolamine by the concerted (enzymatic) activity of many cells. Mutants of M. xanthus selected for resistance to AMI (a mixture of fatty acids) were also resistant to AMV. The possible role of AMV in developmental lysis is discussed.
Antibiotic TA adheres avidly to a variety of surfaces. When administered orally or intravenously ... more Antibiotic TA adheres avidly to a variety of surfaces. When administered orally or intravenously to animals, the antibiotic concentrates in an active form close to the site of administration. No measurable blood level of the antibiotic is found. Tissues treated with antibiotic TA and then washed continuously to remove unbound antibiotic retain bactericidal activity. Antibiotic TA may have applications in circumstances where an active bound antibiotic is desired.
The adhesive antibiotic TA was applied to the dento‐gingival junction of 8 human volunteers, suff... more The adhesive antibiotic TA was applied to the dento‐gingival junction of 8 human volunteers, suffering from moderate to severe gingivitis. 2 diametrically opposed quadrants of the mouth received 4 applications of 0.1 mg TA, while the other 2 quadrants were treated with a placebo and served as controls. The plaque index, gingival index and bleeding index were scored periodically for 2 weeks and in 4 patients for up to 30 days. The TA‐treated quadrants showed a rapid decrease in all 3 indices following 2 treatments. A further improvement was observed with the 3rd and 4th treatments. 30 days after the onset of the experiment and 23 days after the last application, the indices were still considerably lower than the initial values.
The beta-galactosidase activities arising from Tn5lac insertions in several genes required for an... more The beta-galactosidase activities arising from Tn5lac insertions in several genes required for antibiotic TA production were measured under different growth conditions. In all of the non-TA-producing mutants, the beta-galactosidase specific activity was higher when the cells were grown in nutrient-limited 0.5CTS medium (0.5% Casitone plus alanine, serine, and glucose) than in rich 2CT medium (2% Casitone). One of the mutants, 420, had low beta-galactosidase specific activity in both media. The other seven mutants containing inserts in genes essential for TA production had specific activities of 139 to 367 U/mg of protein in 0.5CTS medium and 11 to 48 U/mg of protein in 2CT medium. The beta-galactosidase specific activities of two strains, 1030 and 420, increased during exponential growth in 0.5CTS medium. The beta-galactosidase specific activities of both strains increased greatly when the cells were grown in the presence of magnesium phosphate, which traps ammonium ions. The Tn5lac...
Two bacteriophages for Bdellovibrio bacteriovorus were isolated. One of the phages (VL-1) was iso... more Two bacteriophages for Bdellovibrio bacteriovorus were isolated. One of the phages (VL-1) was isolated on a host-independent Bdellovibrio strain, and the other (VL-2) was isolated on a host-dependent strain. Both phages grew on host-dependent as well as on host-independent Bdellovibrio strains. The development of the phages in host-dependent bdellovibrios occurred only when the phage-infected bdellovibrios parasitized cells of other bacteria. In the absence of other bacteria, the phages adsorbed to the bdellovibrios and killed them and in the process lost their own plaque-forming ability.
Ultrastructure was correlated with growth kinetics of bdellophage VL-1 infecting host-dependent (... more Ultrastructure was correlated with growth kinetics of bdellophage VL-1 infecting host-dependent (“parasitic”) Bdellovibrio bacteriovorus 109J in its Escherichia coli B host (the three-membered system), as well as in the host-independent (“saprophytic”) derivative of the Bdellovibrio. Electron microscope observations showed the arrested growth of the phage-infected bdellovibrios, polar localization of the phage progeny, and stages in their release. Present evidence indicates that bdellophage DNA is derived from both the Bdellovibrio and its host cell.
Quantitative methods were developed for the study of the early stages in the interaction of Bdell... more Quantitative methods were developed for the study of the early stages in the interaction of Bdellovibrio bacteriovorus and host bacteria. Attachment measurements were based on the differential filtration of host and parasite. Invasion was measured by estimation of radioactively labeled Bdellovibrio cells remaining attached to the host cells after mechanical agitation. The kinetics of attachment and the final number of Bdellovibrio cells attached were dependent on the multiplicity of the parasite, the composition and p H of the medium, and the incubation temperature. Inhibitors of Bdellovibrio motility, including chelating agents, NaN 3 , and low p H, all inhibited attachment, as did anaerobiosis. Ultraviolet-killed host cells retained their competence for attachment of Bdellovibrio cells, whereas heat-killed cells lost it. Invasion was selectively inhibited by inhibitors of protein synthesis, such as streptomycin, puromycin, and chloramphenicol. These antibiotics had no effect on at...
A bacterial predator-prey interaction was studied using Bdellovibrio and bioluminescent prey bact... more A bacterial predator-prey interaction was studied using Bdellovibrio and bioluminescent prey bacteria. The attacking bdellovibrio causes decay of bioluminescence, which is correlated with bdellovibrio penetration into the prey. The behavior of the prey and predator populations over time was found to be well described by a Lotka-Volterra model. By using this model, the probability of bdellovibrio penetration after encountering a prey cell was found to be approximately 3.0%. The prey density required to give the bdellovibrios a 50% chance of survival was calculated to be at least 3.0 × 10 6 cells per ml, and the density required for population equilibria was calculated to be about 7 × 10 5 prey bacteria per ml. These values, not generally characteristic of natural habitats, suggest that the existence of Bdellovibrio in nature is limited to special ecological niches.
Antibiotic TA (TA) is a wide-spectrum, bactericidal antibiotic produced by Myxococcus xanthus str... more Antibiotic TA (TA) is a wide-spectrum, bactericidal antibiotic produced by Myxococcus xanthus strain TA. It was previously demonstrated that TA binds tightly to soft tissues while retaining its bactericidal activity in the bound form. The present study was undertaken to investigate TA adsorption to dental hard tissues. Slabs of dental tissues that had been cut from periodontally-involved extracted human teeth were treated with TA and then washed in saline with shaking (saline being replaced every 15 minutes). After 30, 60, and 120 minutes of washing, 45, 39, and 27% of the input TA activities were retained on the slabs, respectively. The tooth-bound TA was released slowly into the aqueous medium in active form. Similar experiments with beta-lactam antibiotics resulted in no significant adhesion to the slabs. The study demonstrates that TA binds tightly to dental tissues while retaining its bactericidal properties.
Bdellovibrios capable of axenic growth grow in a cell-free medium at a rate considerably lower th... more Bdellovibrios capable of axenic growth grow in a cell-free medium at a rate considerably lower than that attainable in a two-membered culture with Escherichia coli. The axenic growth rate may be improved either by adjustment of the osmosity of the medium or by the addition of low concentrations of spermine.
Autocide AMV of Myxococcus xanthus was purified and identified as phosphatidylethanolamine. Alkal... more Autocide AMV of Myxococcus xanthus was purified and identified as phosphatidylethanolamine. Alkaline hydrolysis of AMV yielded a high proportion of mono- and diunsaturated fatty acids. The bactericidal activity of AMV on M. xanthus depended upon the density of target cells: the greater the cell density, the greater the killing by AMV. For example, at 2 U of AMV per ml, 0, 50, and 99% killing was measured with 2 X 10(4), 2 X 10(5), and 2 X 10(7) target cells per ml, respectively. The cell-density-dependent activity of AMV was also observed on solid medium. Studies with model lipid compounds suggest that the inhibitory activity of AMV is due to the fatty acid moiety, released from phosphatidylethanolamine by the concerted (enzymatic) activity of many cells. Mutants of M. xanthus selected for resistance to AMI (a mixture of fatty acids) were also resistant to AMV. The possible role of AMV in developmental lysis is discussed.
Antibiotic TA adheres avidly to a variety of surfaces. When administered orally or intravenously ... more Antibiotic TA adheres avidly to a variety of surfaces. When administered orally or intravenously to animals, the antibiotic concentrates in an active form close to the site of administration. No measurable blood level of the antibiotic is found. Tissues treated with antibiotic TA and then washed continuously to remove unbound antibiotic retain bactericidal activity. Antibiotic TA may have applications in circumstances where an active bound antibiotic is desired.
The adhesive antibiotic TA was applied to the dento‐gingival junction of 8 human volunteers, suff... more The adhesive antibiotic TA was applied to the dento‐gingival junction of 8 human volunteers, suffering from moderate to severe gingivitis. 2 diametrically opposed quadrants of the mouth received 4 applications of 0.1 mg TA, while the other 2 quadrants were treated with a placebo and served as controls. The plaque index, gingival index and bleeding index were scored periodically for 2 weeks and in 4 patients for up to 30 days. The TA‐treated quadrants showed a rapid decrease in all 3 indices following 2 treatments. A further improvement was observed with the 3rd and 4th treatments. 30 days after the onset of the experiment and 23 days after the last application, the indices were still considerably lower than the initial values.
The beta-galactosidase activities arising from Tn5lac insertions in several genes required for an... more The beta-galactosidase activities arising from Tn5lac insertions in several genes required for antibiotic TA production were measured under different growth conditions. In all of the non-TA-producing mutants, the beta-galactosidase specific activity was higher when the cells were grown in nutrient-limited 0.5CTS medium (0.5% Casitone plus alanine, serine, and glucose) than in rich 2CT medium (2% Casitone). One of the mutants, 420, had low beta-galactosidase specific activity in both media. The other seven mutants containing inserts in genes essential for TA production had specific activities of 139 to 367 U/mg of protein in 0.5CTS medium and 11 to 48 U/mg of protein in 2CT medium. The beta-galactosidase specific activities of two strains, 1030 and 420, increased during exponential growth in 0.5CTS medium. The beta-galactosidase specific activities of both strains increased greatly when the cells were grown in the presence of magnesium phosphate, which traps ammonium ions. The Tn5lac...
Two bacteriophages for Bdellovibrio bacteriovorus were isolated. One of the phages (VL-1) was iso... more Two bacteriophages for Bdellovibrio bacteriovorus were isolated. One of the phages (VL-1) was isolated on a host-independent Bdellovibrio strain, and the other (VL-2) was isolated on a host-dependent strain. Both phages grew on host-dependent as well as on host-independent Bdellovibrio strains. The development of the phages in host-dependent bdellovibrios occurred only when the phage-infected bdellovibrios parasitized cells of other bacteria. In the absence of other bacteria, the phages adsorbed to the bdellovibrios and killed them and in the process lost their own plaque-forming ability.
Ultrastructure was correlated with growth kinetics of bdellophage VL-1 infecting host-dependent (... more Ultrastructure was correlated with growth kinetics of bdellophage VL-1 infecting host-dependent (“parasitic”) Bdellovibrio bacteriovorus 109J in its Escherichia coli B host (the three-membered system), as well as in the host-independent (“saprophytic”) derivative of the Bdellovibrio. Electron microscope observations showed the arrested growth of the phage-infected bdellovibrios, polar localization of the phage progeny, and stages in their release. Present evidence indicates that bdellophage DNA is derived from both the Bdellovibrio and its host cell.
Quantitative methods were developed for the study of the early stages in the interaction of Bdell... more Quantitative methods were developed for the study of the early stages in the interaction of Bdellovibrio bacteriovorus and host bacteria. Attachment measurements were based on the differential filtration of host and parasite. Invasion was measured by estimation of radioactively labeled Bdellovibrio cells remaining attached to the host cells after mechanical agitation. The kinetics of attachment and the final number of Bdellovibrio cells attached were dependent on the multiplicity of the parasite, the composition and p H of the medium, and the incubation temperature. Inhibitors of Bdellovibrio motility, including chelating agents, NaN 3 , and low p H, all inhibited attachment, as did anaerobiosis. Ultraviolet-killed host cells retained their competence for attachment of Bdellovibrio cells, whereas heat-killed cells lost it. Invasion was selectively inhibited by inhibitors of protein synthesis, such as streptomycin, puromycin, and chloramphenicol. These antibiotics had no effect on at...
A bacterial predator-prey interaction was studied using Bdellovibrio and bioluminescent prey bact... more A bacterial predator-prey interaction was studied using Bdellovibrio and bioluminescent prey bacteria. The attacking bdellovibrio causes decay of bioluminescence, which is correlated with bdellovibrio penetration into the prey. The behavior of the prey and predator populations over time was found to be well described by a Lotka-Volterra model. By using this model, the probability of bdellovibrio penetration after encountering a prey cell was found to be approximately 3.0%. The prey density required to give the bdellovibrios a 50% chance of survival was calculated to be at least 3.0 × 10 6 cells per ml, and the density required for population equilibria was calculated to be about 7 × 10 5 prey bacteria per ml. These values, not generally characteristic of natural habitats, suggest that the existence of Bdellovibrio in nature is limited to special ecological niches.
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